Pyrroloindole derivatives and intermediates in producing the same

ABSTRACT

PCT No. PCT/JP98/00234 Sec. 371 Date Jul. 19, 1999 Sec. 102(e) Date Jul. 19, 1999 PCT Filed Jan. 22, 1998 PCT Pub. No. WO98/32757 PCT Pub. Date Jul. 30, 1998Pyrroloindole derivatives having antimicrobial and antitumor activities and having a carbamoyloxy group represented by the following general formula (1), optical isomers thereof, and pharmacologically acceptable salts thereof; and intermediates for production thereof: where R1 is OH, or a pyrrolidinyl group; n is 1 or 2; R2 is a lower alkyl group of C1-C4; X-Y or Y-X is CH2, CHOH, CH2-CH2, O-CH2, or NMe-CH2; Z1 is Cl or Br; and Ar1 is wherein Z2 and Z3 are O or NH; m is 0 or an integer of 1 to 4; and Ar2 is any of the above groups a, b, c, and d.

TECHNICAL FIELD

The present invention relates to novel pyrroloindole derivatives havinga carbamoyloxy group which have antimicrobial and antitumor activities,optical isomers thereof, and pharmacologically acceptable salts thereof;and intermediates for production thereof.

BACKGROUND ART

CC-1065, which is an antibiotic having antimicrobial activity andantitumor activity, is disclosed in J.Antibiotics, vol.31, p.1211(1978), and vol.34, p.1119 (1981); and U.S. Pat. No. 4,169,888.Duocarmycin A having analogous structure, and analogues thereof aredisclosed in WO87/06265; EP0318056; J.Antibiotics, vol.42, p.1229(1989); and JP-A-4-99774. Derivatives of CC-1065 are disclosed inEP0359454, JP-A-60-193989, and published Japanese translation of PCTapplication (Kohyo) 2-502005. Derivatives of duocarmycins are disclosedin JP-A-3-7287, JP-A-3-128379, EP0354583, and EP0406749. These arederived by utilizing directly a basic skeleton of a natural substance,or modification of a natural product. Further, a chemically synthesizedpyrroloindole derivative having a trifluoromethyl group on thepyrroloindole ring is disclosed by the inventors of the presentinvention in JP-A-6-116269.

The inventors of the present invention made comprehensive researches onpyrroloindole derivatives which are effective against solid tumor and isless toxic. Consequently, The inventors found a derivative having atrifluoromethyl group in the pyrroloindole ring, and disclosed it inJP-A-6-116269. In JP-A-6-116269, a prodrug represented by Formula Ahaving a carbamoyl group containing a cyclic amine is disclosed.##STR3##

The present invention intends to provide a prodrug which forms an activespecies (a compound of Formula A in which B is hydrogen) in vivo morereadily than the prodrug shown in Examples of JP-A-6-116269, and iseffective against solid tumor and is less toxic.

DISCLOSURE OF INVENTION

It was found by the inventors of the present invention that thepyrroloindole derivatives having a carbamoyloxy group represented by thefollowing general formula (1) below, optically active isomers thereof,and pharmacologically acceptable salts thereof are useful as theaforementioned prodrug. ##STR4## where R¹ is OH, or a pyrrolidinylgroup; n is 1 or 2; R² is a lower alkyl group of C₁ -C₄ ; X--Y or Y--Xis CH₂, CHOH, CH₂ -CH₂, O--CH₂, or NMe--CH₂ ; Z¹ is Cl or Br; and Ar¹ is##STR5## (wherein Z² is O or NH, m is 0 or an integer of 1 to 4),##STR6## (wherein m is 0 or an integer of 1 to 4), ##STR7## (wherein mis 0 or an integer of 1 to 4), ##STR8## (wherein Z³ is O or NH; Ar² isany of the above groups a, b, c, and d). Consequently the presentinvention has been completed.

The present invention further relates to protected pyrroloindolederivatives represented by the following general formula (2): ##STR9##where R¹ is OR³ (R³ is a protecting group for the hydroxyl group); n is1 or 2; R² is a lower alkyl of C₁ -C₄ ; X--Y is CH₂, CHOH, CH₂ --CH₂,O--CH₂, or NMe--CH₂ ; Z¹ is Cl or Br; and Ar¹ is ##STR10## (wherein Z²is O or NH, m is 0 or an integer of 1 to 4), ##STR11## (wherein m is 0or an integer of 1 to 4), ##STR12## (wherein m is 0 or an integer of 1to 4), ##STR13## (wherein Z³ is O or NH; Ar³ is any of the above groupsof a, b, c, and d), and further relates to protected pyrroloindolederivatives represented by the following general formula (2): ##STR14##where R¹ is OR³ (R³ is a protecting group for the hydroxyl group), n is1, R² is methyl, X--Y is O--CH₂, Z¹ is Cl, and Ar¹ is ##STR15##

These derivatives are useful as an intermediate for production of theaforementioned carbamoyl group-containing pyrroloindole derivative.

The present invention further relates to an intermediate for productionof the aforementioned protected pyrroloindole derivative represented bythe following general formula (3), optically active isomers thereof, andsalts thereof: ##STR16## where R³ is a protecting group for the hydroxylgroup, R⁴ is a hydrogen atom or a benzyl group.

In the general formulas (1) and (2), the lower alkyl group of C₁ -C₄includes methyl, ethyl, isopropyl, t-butyl, and the like, of whichmethyl is preferred. The hydroxyl-protecting group represented by R³includes t-butyldimethylsilyl, triethylsilyl, triisopropylsilyl,t-butyldiphenylsilyl, methoxymethyl, t-butoxymethyl, tetrahydropyranyl,and the like, of which t-butyldimethylsilyl is preferred.

In the present invention, particularly preferred embodiment isrepresented by the following general formula (4) below: ##STR17##

Of the compounds, preferred are the compound in which R¹ ispyrrolidinyl, and X--Y is CH₂ (Compound I); the compound in which R₁ isOH, and X--Y is CH₂ --CH₂ (Compound II); the compound in which R¹ is OH,and X--Y or Y--X is O--CH₂ (Compound III); and the compound in which R¹is OH, and X--Y or Y--X is NMe--CH₂ (Compound IV); and optically activeisomers of the above compounds. The compounds I-IV and the opticallyactive isomers thereof may be pharmacologically accepted salts, forexample, a hydrochloride salt.

The racemic modifications and optically active isomers of the compoundsrepresented by the general formula (1) can be produced by a knownprocess (e.g., JP-A-6-116269). The compound represented by the generalformula (1) may be used alone or in combination with a pharmaceuticallyacceptable auxiliary agent as an antimicrobial or antitumor composition.

For example, the compound represented by the general formula (1) or thesalt thereof is dissolved in physiological saline or an aqueous solutionof glucose, mannitol, lactose, or the like for use as a medicalcomposition.

Otherwise, a salt of the compound represented by the general formula (1)is freeze-dried in a conventional manner and mixed with sodium chlorideor the like for use as a powdery injectable agent. This medicalcomposition may contain an additive known in medical formulation, forexample, a pharmaceutically acceptable salt, as necessary.

The solution-type medical composition can be used as it is. The powderyinjectable agent is used after dissolution in distilled water,physiological saline, or an aqueous solution of glucose, mannitol,lactose, or the like for intravenous administration. The medicalcomposition may be administered intra-arterially, intra-abdominally, orintrathoracically, if desired.

The medical composition may be used also for oral administration astablets, capsules, powders, granules, ampule-medicine, and so forth, andmay contain a medical auxiliary agent conventionally employed in medicalformulation.

The dosage depends on the age and the symptom of the patient, rangingfrom 0.00001 to 100 mg/kg/day for mammals including humans. Theadministration may be conducted once or several times per day, orintermittently one to four times per week, or once in two to four weeks.

BEST MODE FOR CARRYING OUT THE INVENTION

The advantages of the present invention are described below by referenceto Test Examples and Working Examples without limiting the inventionthereto.

WORKING EXAMPLE 1 ##STR18##

Methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(10.2 mg, 15 μmol) and 4-nitrophenyl chloroformate (7.6 mg, 38 μmol)were dissolved in 1 mL of tetrahydrofuran, and thereto triethylamine(4.2 μL, 30 μmol) was added dropwise with ice cooling. After one hour,(2S)-pyrrolidine-2-methanol (4.4 μL, 45 μmol) was added dropwise. Themixture was stirred for two hours, and stirred further for two hours atroom temperature. The reaction mixture was poured into 3 mL of saturatedaqueous sodium chloride solution and 3 mL of ethyl acetate forextraction. The extracted matter was dried over anhydrous sodiumsulfate, and the solvent was evaporated off. The resulting residue waspurified by silica gel column chromatography (hexane:acetone=1:1) toobtain 8.5 mg (70%) of methyl(1S)-chloromethyl-5-{[(2S)-hydroxymethylpyrrolidin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate.

NMR(DMSO-d₆) 67 : 1.80-2.10 (4 H,m), 3.30-3.80 (6 H,m), 3.83-4.20 (2H,m), 3.92 (3 H,s), 4.01 (3 H,s), 4.42 (1 H,brs), 4.60 (1 H,d, J=9.5Hz), 4.79 (1 H,t, J=9.3 Hz), 7.10 (1 H,dd, J=1.0 Hz,8.1 Hz), 7.21 (1H,d, J=1.5 Hz), 7.28 (1 H,t, J=7.8 Hz), 7.37 (1 H,dd, J=1.0 Hz,7.8 Hz),7.50 (1 H,d, J=8.8 Hz), 7.60 (1 H,dd, J=2.0 Hz, 9.0 Hz), 7.75 (1 H,s),8.17 (1 H,d, J=10.3 Hz), 8.21 (1 H,d, J=1.5 Hz), 10.41 (1 H,s), 11.70 (1H,s), 13.06 (1 H,brs)

Working Example 2 ##STR19##

In the same manner as in Working Example 1, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(20.4 mg, 30 μmol) and (2S,4R)-4-hydroxypyrrolidin-2-methanol (10.5 mg,90 μmol) were allowed to react to obtain methyl (1S)-chloromethyl-5-{[(4R)-hydroxy-(2S)-hydroxymethylpyrrolidin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(17.3 mg, 70%).

NMR(DMSO-d₆) 67 : 1.80-2.20 (2 H,m) , 3.34-3.80 (6 H,m) , 3.90-4.35 (2H,m), 3.92 (3 H,s), 4.01 (3 H,s), 4.35-4.50 (2 H,m), 4.60 (1 H,d, J=11.2Hz), 4.75-4.81 (1 H,m), 5.08-5.15 (1 H,m), 7.10 (1 H,d, J=8.3 Hz), 7.21(1 H,d, J=1.5 Hz), 7.28 (1 H,t, J=7.8 Hz), 7.37 (1 H, d=7.8Hz), 7.50 (1H,d, J=8.8 Hz), 7.60 (1 H,dd, J=1.5 Hz,8.8 Hz), 7.75 (1 H,s), 8.18-8.30(2 H,m), 10.41 (1 H,s), 11.71 (1 H,s), 13.09 (1 H,brs)

Working Example 3 ##STR20##

In the same manner as in Working Example 1, methyl(1S)-chloromethy-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(20.4 mg, 30 μmol) and (2S)-(pyrrolidin-1-ylmethyl)pyrrolidine (13.9 mg,90 μmol) were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(2S)-(pyrrolidin-l-ylmethyl)pyrrolidin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate.The resulting reaction product was treated with 3M-HCl in methanol(0.5mL)-ethyl acetate(0.05 mL) to obtain 18.2 mg (68%) of the hydrochloride.

NMR(DMSO-d₆) δ: 1.80-2.30 (8 H,m), 2.95-4.05 (10 H,m), 3.91 (3 H,s),4.00 (3 H,s), 4.28 (1 H,m), 4.41 (1 H,m), 4.61 (1 H,m), 4.80 (1 H,m),7.10 (1 H,d, J=8.3 Hz), 7.22 (1 H,s), 7.28 (1 H,t, J=7.8 Hz), 7.37 (1H,d, J=7.8 Hz), 7.51 (1 H,d, J=8.8 Hz), 7.61 (1 H,dd, J=1.5 Hz, 8.8 Hz),7.76 (1 H,s), 8.19-8.31 (2 H,m), 9.71 (1 H,brs), 10.43 (1 H,s), 11.67 (1H,s), 13.20 (1 H,brs)

Working Example 4 ##STR21##

In the same manner as in Working Example 1, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(13.6 mg, 20 μmol) and (2RS)-piperidine-2-methanol (6.9 mg, 60 μmol)were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(2RS)-hydroxymethylpiperidin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(12.5 mg, 76%).

NMR(DMSO-d₆) )δ: 1.40-1.85 (6 H,m), 3.25-3.95 (8 H,m), 3.91 (3 H,s),4.00 (3 H,s), 4.42 (1 H,m), 4.60 (1 H,d, J=10.7 Hz), 4.80 (1 H,t, J=9.8Hz), 7.10 (1 H,d, J=8.3 Hz), 7.21 (1 H,s), 7.28 (1 H,t, J=7.8 Hz), 7.36(1 H,d, J=7.8 Hz), 7.49 (1 H,d, J=8.8 Hz), 7.60 (1 H,dd, J=2.0 Hz, 9.3Hz), 7.75 (1 H,s), 8.21 (2 H,m), 10.41 (1 H,s), 11.74 (1 H,s)

Working Example 5 ##STR22##

In the same manner as in Working Example 3, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(27.2 mg, 40 μmol) and (2RS)-4-methylpiperazine-2-methanol (13.0 mg, 100μmol) were allowed to react and the resulting product was treated toobtain hydrochloride of methyl(1S)-chloromethyl-5-{[(2RS)-hydroxymethyl-4-methylpiperazin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(18.8 mg, 54%).

NMR(DMSO-d₆) δ: 2.88 (3 H,s) , 3.05-3.95 (10 H,m), 3.92 (3 H,s), 4.00 (3H,s), 4.14 (1 H,m), 4.20-4.50 (2 H,m), 4.61 (1 H,d, J=11.2 Hz), 4.81 (1H,t, J=9.3 Hz), 7.10 (1 H,d, J=7.8 Hz), 7.22 (1 H,s), 7.28 (1 H,t, J=7.8Hz), 7.36 (1 H,d, J=7.8 Hz), 7.50 (1 H,d, J=9.3 Hz), 7.61 (1 H,d, J=10.3Hz), 7.76 (1 H,s), 8.21-8.31 (2 H,m), 10.00 (1 H,br), 10.43 (1 H,s),11.66 (1 H,br), 13.11 (1 H,brs)

Working Example 6 ##STR23##

In the same manner as in Working Example 3, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(20.4 mg, 30 μmol) and (2RS)-4-methylpiperazine-2-ethanol (12.4 mg, 90μmol) were allowed to react and the resulting product was treated toobtain hydrochloride of methyl(1S)-chloromethyl-5-{[(2RS)-hydroxyethyl-4-methylpiperazin-1-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(16.5 mg, 62%).

NMR(DMSO-D₆) )δ: 1.80-2.50 (2 H,m), 2.86 (3 H,s), 3.05-3.95 (10 H,m),3.92 (3 H,s), 4.00 (3 H,s), 4.08 (1 H,m), 4.43 (1 H,m), 4.59 (1 H,m),4.80 (1 H,m), 4.94 (1 H,m), 7.10(1 H,d, J=7.8 Hz), 7.22 (1 H,d, J=2.0Hz), 7.28 (1 H,t, J=7.8 Hz), 7.37 (1 H,dd, J=1.0 Hz, 8.8 Hz), 7.50 (1H,d, J=8.8 Hz), 7.61 (1 H,dd, J=2.0 Hz, 8.8 Hz), 7.76 (1 H,s), 8.21-8.32(2 H, m), 10.05 (1 H,brs), 10.43 (1 H,s), 11.67 (1 H,s), 13.17 (1 H,brs)

Working Example 7 ##STR24##

In the same manner as in Working Example 1, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(20.4 mg, 30 μmol) and (3RS)-morpholine-3-methanol (10.5 mg, 90 μmol)were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(3RS)-hydroxymethylmorpholin-4-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(17.5 mg, 71%).

NMR(DMSO-d₆) 67 : 3.15-4.05 (11 H,m), 3.92 (3 H,s), 4.00 (3 H,s), 4.29(1 H,m), 4.43 (1 H,m), 4.61 (1 H,d, J=10.7Hz), 4.80 (1 H,t, J=9.3 Hz),7.10 (1 H,d,J=7.3 Hz), 7.22 (1 H,d, J=2.0 Hz), 7.28 (1 H,t, J=8.3 Hz),7.36 (1 H,d, J=7.8 Hz), 7.49 (1 H,d, J=8.8 Hz), 7.60 (1 H,dd, J=2.0Hz,8.8 Hz), 7.75 (1 H,s), 8.21 (2 H,m), 10.41 (1 H,s), 11.73 (1 H,s),13.08 (1 H,brs)

Working Example 8 ##STR25##

In the same manner as in Working Example 1, methyl(1S)-chloromethyl-5-hydroxy-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(81.7 mg, 0.12 μmol) and (3R)-morpholine-3-methanol (42.2 mg, 0.36 mmol)were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(3R)-hydroxymethylmorpholin-4-ylcarbonyl]oxy}-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate (62.9 mg, 64%).

NMR(DMSO-d₆) δ: 3.15-4.15 (11 H,m), 3.91 (3 H,s), 4.00 (3 H,s), 4.30 (1H,m), 4.42 (1 H,m), 4.60 (1 H,d, J=10.7 Hz), 4.80 (1 H,t, J=10.3 Hz),7.10 (1 H,d, J=7.3 Hz), 7.22 (1 H,s), 7.28 (1 H,t, J=7.8 Hz), 7.37 (1H,d, J=7.8 Hz), 7.49 (1 H,d, J=8.5 Hz), 7.60 (1 H,dd, J=2.0 Hz, 9.0 Hz),7.76 (1 H,s), 8.21 (2 H,s), 10.42 (1 H,s), 11.74 (1 H,brs), 12.78,13.09(total 1 H,brsx2, in each rotamer)

Working Example 9 ##STR26##

In the same manner as in Working Example 1, methyl(1S)-chloromethyl-5-hydroxy-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(81.7 mg, 0.12 mmol) and (3S)-morpholine-3-methanol (42.2 mg, 0.36 mmol)were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(3S)-hydroxymethylmorpholin-4-ylcarbonyl]oxy}-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate (69.7 mg, 70%).

NMR(DMSO-d₆) δ: 3.10-4.15 (11 H,m), 3.91 (3 H,s), 4.00 (3 H,s), 4.28 (1H,m), 4.43 (1 H,m), 4.60 (1 H,d, J=11.0 Hz), 4.80 (1 H,t, J=10.5 Hz),7.10 (1 H,d, J=7.3 Hz), 7.23 (1 H,d, J=2.0 Hz), 7.28 (1 H,t, J=7.8 Hz),7.37 (1 H,d, J=7.8 Hz), 7.49 (1 H,d, J=9.0 Hz), 7.60 (1 H,dd, J=2.0 Hz,8.8 Hz), 7.76 (1 H,s), 8.21 (2 H,m), 10.42 (1 H,s), 11.73 (1 H,s),12.85, 13.11 (total 1 H,brsx2, in each rotamer).

Working Example 10 ##STR27##

Methyl(1S)-chloromethyl-5-hydroxy-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.02 g, 1.5 mmol) and 4-nitrophenyl chloroformate (0.51 g, 2.55 mmol)were dissolved in 50 mL of tetrahydrofuran, and thereto triethylamine(313.6 μL, 2.25 mmol) was added dropwise with ice cooling. After 1.5hours, (3S)-3-(t-butyldimethylsilyloxy)-methylmorpholine hydrochloride(0.80 g, 3.0 mmol) and triethylamine (0.52 mL, 3.75 mmol) were addedthereto. The mixture was stirred overnight. The reaction mixture wasdiluted with 50 mL of ethyl acetate, washed with saturated aqueoussodium chloride solution, and dried over anhydrous sodium sulfate. Afterthe solvent was evaporated off, the resulting residue was purified bysilica gel column chromatography (hexane:ethyl accetate=1:1) to obtainmethyl(1S)-chloromethyl-5-{[(3S)-(t-butyldimethylsilyloxy)-methylmorpholin-4-ylcarbonyl]oxy}-3-[5-[7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.08 g, 77%).

NMR(DMSO-d₆) )δ: 0.04,0.07 (total 6 H,sx2, in each rotamer), 0.85,0.86(total 9H,sx2, in each rotamer), 3.15-4.10 (10 H,m), 3.91 (3 H,s), 4.00(3 H,s), 4.19 (1 H,m), 4.40 (1 H,m), 4.59 (1 H,d, J=11.2 Hz), 4.80 (1H,t, J=10.7 Hz), 7.10 (1 H,d, J=7.8 Hz), 7.22 (1 H,s), 7.28 (1 H,t,J=8.1 Hz), 7.37 (1 H,d, J=7.6 Hz), 7.50 (1 H,d, J=8.8 Hz), 7.60 (1 H,dd,J=2.0 Hz,9.0 Hz), 7.76 (1 H,s), 8.17 (2 H,m), 10.42 (1 H,s), 11.69 (1H,s), 13.09 (1 H,brs)

Working Example 11 ##STR28##

In the same manner as in Working Example 10, methyl(1S)-chloromethyl-5-hydroxy-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.36 g, 2.0 mmol) and (3R)-3-(t-butyldimethylsilyloxy)-methylmorpholinehydrochloride (1.07 g, 4.0 mmol) were allowed to react to obtain methyl(1S)-chloromethyl-5-{[(3R)-(t-butyldimethylsilyloxy)-methylmorpholin-4-ylcarbonyl]oxy]-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.58 g, 84%).

NMR (DMSO-d₆) δ: 0.07,0.08 (total 6 H,sx2, in each rotamer), 0.867,0.874(total 9 H,sx2, in each rotamer), 3.15-4.10 (10 H,m), 3.91 (3 H,s), 4.00(3 H,s), 4.21 (1 H,m), 4.41 (1 H,m), 4.59 (1 H,d,J=10.5 Hz), 4.80 (1H,t,J=10.2 Hz), 7.10 (1 H,d,J=7.6 Hz), 7.22 (1 H,s), 7.28 (1 H,t,J=7.8Hz), 7.37 (1 H,d,J=7.8 Hz), 7.50 (1 H,d,J=8.8 Hz), 7.60 (1 H,dd,J=2.0Hz,8.8 Hz), 7.76 (1 H,s), 8.18 (1 H,s), 8.20 (1 H,s), 10.42 (1 H,s),11.69 (1 H,s), 13.08 (1 H,brs)

Working Example 12 ##STR29##

Methyl(1S)-chloromethyl-5-[(3S)-(t-butyldimethylsilyloxy)methylmorpholin-4-ylcarbonyl)oxy]-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.03 g, 1.1 mmol) was suspended in 2 mL of ethyl acetate and 8 mL ofisopropanol. Thereto, 5.5 mL of 1 N hydrochloric acid in isopropylalcohol was added, and the mixture was stirred at room temperature forone hour. The reaction mixture was cooled by ice. Thereto, 10 mL ofisopropyl ether was added and the mixture was stirred for 15 minutes.The formed crystalline matter was collected by filtration, and washedwith isopropyl ether to obtain methyl(1S)-chloromethyl-5-{[(3R)-hydroxymethylmorpholin-4-ylcarbonyl]oxy}-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(0.83 g, 92%) as colorless crystals. This substance is the same as theone obtained in Working Example 8.

Working Example 13 ##STR30##

In the same manner as in Working Example 12, methyl(1S)-chloromethyl-5-{[(3R)-(t-butyldimethylsilyloxy)-methylmorpholin-4-ylcarbonyl]oxy}-3-[5-[(7-methoxybenzofuran-2-ylcarbonyl)amino]-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.41 g, 1.5 mmol) was treated to obtain methyl(1S)-chloromethyl-5-{[(3S)-hydroxymethylmorpholin-4-ylcarbonyl]oxy}-3-[5-(7-methoxybenzofuran-2-ylcarbonyl)amino-1H-indol-2-ylcarbonyl]-7-trifluoromethyl-1,2,3,6-tetrahydropyrrolo[3,2-e]indole-8-carboxylate(1.20 g, 97%) as colorless crystals. This substance is the same as theone obtained in Working Example 9.

Working Example 14 ##STR31##

(3R)-4-Benzylmorpholine-3-methanol (1.04 g, 5.0 mmol),t-butyldimethylchlorosilane (0.83 g, 5.5 mol), and imidazole (0.41 g,6.0 mmol) were allowed to react in 5 mL of dichloromethane at roomtemperature overnight. The reaction product was purified by silica gelcolumn chromatography (hexane:ethyl acetate=5:1) to obtain(3S)-3-(t-butyldimethylsilyloxy)methyl-4-benzylmorpholine (1.53 g, 95%)as a colorless oil.

Working Example 15 ##STR32##

In the same manner as in Working Example 14,(3S)-4-benzylmorpholine-3-methanol (1.24 g, 6.0 mmol) was treated toobtain (3R)-3-(t-butyldimethylsilyloxy)methyl-4-benzylmorpholine (1.82g, 94%) as a colorless oil.

Working Example 16 ##STR33##

(3R)-Morpholine-3-methanol (84.8 mg, 0.72 mmol),t-butyldimethylchlorosilane (0.11 g, 0.72 mmol), and imidazole (59.1 mg,0.87 mmol) were allowed to react in 1 mL of dichloromethane at roomtemperature for 2 hours. The reaction product was purified by silica gelchromatography (dichloromethane:methanol:acetone=15:1:0.5) to obtain(3S)-3-(t-butyldimethylsilyloxy)methylmorpholine (0.15 g, 92%) as acolorless oil.

Working Example 17 ##STR34##

(3S)-3-(t-Butyldimethylsilyloxy)methyl-4-benzylmorpholine (1.19 g, 3.7mmol) was treated for hydrogenation in the presence of 10%palladium-carbon (0.36 g) in 5 mL of ethanol and 0.32 mL of chloroformfor 4 hours (one atmosphere). The catalyst was removed by filtration,and the filtrate was concentrated. The precipitated crystalline matterwas washed with ether to obtain(3S)-3-(t-butyldimethylsilyloxy)methylmorpholine hydrochloride (0.96 g(96%).

Working Example 18 ##STR35##

In the same manner as in working Example 17,(3R)-3-(t-butyldimethylsilyloxy)methyl-4-benzylmorpholine (1.82 g, 5.6mmol) was treated to obtain(3R)-3-(t-butyldimethylsilyloxy)methylmorpholine hydrochloride (1.13 g,74%).

Test Example 1

Decomposition reaction under chemical conditions:

A buffer solution of pH 7.4 was prepared by mixing suitably an aqueous0.1 M citric acid solution and an aqueous 0.2 M disodiumhydrogenphosphate solution. The buffer solution was diluted tenfold withwater. This buffer solution was further dissolved inN,N-dimethylacetamide at a concentration of 50 v/v%, and its pH wasadjusted to 7.4 by adding suitable amount of aqueous 0.1 M citric acidsolution to prepare a buffer solution. The test compounds were dissolvedrespectively in the buffer solution at a concentration of 5 μM toprepare the test sample solutions. The test sample solutions were placedrespectively in a glass vial, and the glass vials were kept at 37° C. ina thermostat. The change of the concentration of remaining testcompounds with lapse of time was determined quantitativelyintermittently by high performance liquid chromatography (HPLC) tomeasure the half life (t_(1/2) : hours). Table 1 shows the test results.

                  TABLE 1                                                         ______________________________________                                        Test     Half life     Test     Half life                                       compound (t.sub.1/2 : hours) compound (t.sub.1/2 : hours)                   ______________________________________                                        Example 1                                                                              >8            Example 5                                                                              1.7*                                            Example 2 >8 Example 6 5.8*                                                   Example 3   1.0 Example 7 1.7*                                                Example 4  >8*                                                              ______________________________________                                         *Average of the half lives of the diastereomers                          

The compounds of the present invention decomposes rapidly in the buffersolution of pH 7.4 to form the active species.

Test Example 2

Evaluation with mouse bearing M5076/ADR cells (adriamycin-resistantstrain) transplanted subcutaneously:

M5076/ADR cells (adriamycin-resistant strain) were transplantedsubcutaneously at the axillary region of female mice (BDFI strain, 8week age) in an amount of 3.0×10⁶ cells for an individual mouse. Ninedays after the transplantation, the test compound solution was injectedonce into the tail vein. After 13 days from the injection, the tumor wasexcised and weighed. The antitumor activity was evaluated by the ratio(T/C) of the average tumor weight (T) of the administrated group to theaverage tumor weight (C) of control group to which solvent wasadministrated. Table 2 shows the effects.

                  TABLE 2                                                         ______________________________________                                        Test compound   Dose (mg/kg)                                                                             T/C                                                ______________________________________                                        Example 4       1.0        0.01                                                 Example 5 1.0 0.01                                                            Example 6 1.0 0.02                                                          ______________________________________                                    

The compounds of the present invention showed excellent antitumor effectagainst M5076/ADR cells (adriamycin-resistant strain).

INDUSTRIAL APPLICABILITY

The pyrroloindole derivative of the present invention, which has acarbamoyloxy group comprising a cyclic amine having hydroxyalkyl orpyrrolidinylalkyl at α-position to the nitrogen of the carbamoyl group,decomposes at an appropriate rate in vivo or under chemical conditionsto produce active species, although carbamoyl groups comprising a cyclicamine are generally st able. Such a prodrug have firstly producedaccording to the present invention. The prodrug compounds of the presentinvention are effective against solid tumor, and are less toxic andexhibit antitumor activity in a broad safety range. Therefore thechemotherapy for patients with cancer can be conducted with less adverseeffects.

What is claimed is:
 1. A pyrroloindole derivative having a carbamoyloxygroup represented by the following general formula (1) below, anoptically active isomer thereof, and a pharmacologically acceptable saltthereof: ##STR36## where R¹ is OH, or a pyrrolidinyl group; n is 1 or 2;R² is a lower alkyl group of C₁ -C₄ ; X--Y or Y--X is CH₂, CHOH, CH₂-CH₂, O--CH₂, or NMe--CH₂ ; Z¹ is Cl or Br; and Ar¹ is ##STR37## whereinZ² is O or NH, m is 0 or an integer of 1 to 4, ##STR38## wherein m is 0or an integer of 1 to 4, ##STR39## wherein m is 0 or an integer of 1 to4, ##STR40## wherein Z³ is O or NH; Ar² is any of the above groups a, b,c, and d.
 2. The pyrroloindole derivative having a carbamoyloxy groupaccording to claim 1 represented by the following general formula (1)below, the optically active isomer thereof, and the pharmacologicallyacceptable salt thereof: ##STR41## where R¹ is a pyrrolidinyl group, nis 1, R² is methyl, X--Y is CH₂, Z¹ is Cl, and Ar¹ is ##STR42##
 3. Thepyrroloindole derivative having a carbamoyloxy group according to claim1 represented by the following general formula (1) below, and theoptically active isomer thereof: where R¹ is OH, n is 1, R² is methyl,X--Y is CH₂ -CH₂, Z¹ is Cl, and Ar¹ is ##STR43##
 4. The pyrroloindolederivative having a carbamoyloxy group according to claim 1 representedby the following general formula (1) below and the optically activeisomer thereof: where R¹ is OH, n is 1, R² is methyl, X--Y or Y--X isO--CH₂, Z¹ is ##STR44##
 5. The pyrroloindole derivative having acarbamoyloxy group according to claim 1 represented by the followinggeneral formula (1) below, the optically active isomer thereof, and thepharmacologically acceptable salt thereof: where R¹ is OH, n is 1, R² ismethyl, X--Y or Y--X is NMe--CH₂, Z¹ is Cl, and Ar¹ is ##STR45##
 6. Aprotected pyrroloindole derivative represented by the following generalformula (2): where R¹ is OR³ wherein R³ is a protecting group for thehydroxyl group; n is 1 or 2; R² is a lower alkyl of C₁ -C₄ ; X--Y isCH₂, CHOH, CH₂ -CH₂, O--CH₂, or NMe--CH₂ ; Z¹ is Cl or Br; and Ar¹ is##STR46## wherein Z² is O or NH, m is 0 or an integer of 1 to 4,##STR47## wherein m is 0 or an integer of 1 to 4, ##STR48## wherein m is0 or an integer of 1 to 4, ##STR49## wherein Z³ is O or NH; Ar² is anyof the above groups of a, b, c, and d).
 7. The protected pyrroloindolederivative according to claim 6 represented by the following generalformula (2): ##STR50## where R¹ is OR³ wherein R³ is a protecting groupfor the hydroxyl group, n is 1, R² is methyl, X--Y is O--CH₂, Z¹ is Cl,and Ar¹ is ##STR51##